IMPETIGO HUMEDO Y SECO PDF

libros de cátedra introducción la microbiología clínica horacio lopardo ( coordinador) facultad de ciencias exactas introducció la micr robiolo ogía línica. 20 Set. List of contexts by syntactic structures. On words · On functions. On words. Lemma. –, a, a ab abs, a latere, a nativitate Domini, Aaron, ab. seco, trabajo húmedo, y productos de limpieza estan asociadas al riesgo de reportar He called “impetigo sparsa” to eruptions in the.

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Albumin fusion proteins and beta interferon. The invention relates generally to therapeutic proteins including, among others, at least one polypeptide, antibody, peptide or fragment and variant thereof fused to albumin or fragments or variants of albumin 5. The invention encompasses polynucleotides encoding therapeutic proteins albumin fusion, therapeutic albumin fusion proteins, composition, pharmaceutical compositions and kits.

At present, HA for clinical use is produced by extraction from human blood. Therapeutic in its native state or when recombinantly produced, such as interferons and growth hormones form proteins are typically labile molecules exhibiting short half-lives, particularly when formulated in aqueous solutions.

Few practices have been proposed to storage problems of labile protein molecules solutions. According to the above, a need exists for long acting formulations stabilized proteinaceous therapeutic molecules that are easily dispensed, preferably with a simple formulation requiring minimal post 25 conservation manipulation. The present invention provides an expression vector encoding the leader sequence of IFN-beta native 30 fused to a fusion protein of albumin IFN-beta having the mature condensed-beta IFN with the amino form of the protein of the form mature HSA, wherein the expression vector is the expression vector pEE12,1 NS0.

The present invention also provides a mammalian host cell comprising the expression vector of the invention. The present invention also provides a 35 process for producing a fusion protein of albumin which comprises cultivating a host cell of the invention under conditions suitable for expression of the fusion protein of albumin and isolating the fusion protein of the conditions albumin.

Herein pharmaceutical formulations comprising a fusion protein of albumin of the invention and a pharmaceutically acceptable diluent or carrier are also disclosed.

Dichas formulaciones pueden 55 estar en un kit o recipiente. Such formulations may be in kit 55 or container. Such kit or container may be packaged with instructions pertaining to the extended impeitgo of the therapeutic protein.

Dichas formulaciones se pueden usar en procedimientos de Such formulations may be used in methods of.

This document procedures for prevention, treatment or amelioration of a disease or disorder are also disclosed. Y” de la Tabla 1 en una cantidad eficaz para tratar, prevenir o mejorar la enfermedad o trastorno. In preferred embodiments, the present invention encompasses fusion proteins albumin 5 for use in treating a disease or listed in column disorder “Preferred Indication: Y” of Table 1 comprising administering to a patient in which desired such treatment, prevention or amelioration a fusion protein of albumin of the invention comprising a therapeutic protein or a portion corresponding to a therapeutic kmpetigo or fragment or variant thereof disclosed in a column “therapeutic protein: X” Table 1 in the same row as the disease or disorder to be treated is indicated at 10 “preferred Indication: Y” column of Table 1 in an amount effective to treat, prevent or ameliorate the disease or disorder.

The fusion protein of albumin described in Table 1 or 2 has an extended shelf life. This document also disclosed transgenic organisms modified to contain nucleic acid molecules of the invention the polynucleotides described in Tables 1 and 2preferably modified to express a fusion protein of albumin of the invention. After 24 hours the supernatants 25 indicator cells were extracted and analyzed SEAP activity.

The fusion protein of albumin and IFNb was purified from three stable clones. IFNb mammalian derived, Avonex, Biogen came from and was reported to have a specific activity of 2. The following definitions are provided to facilitate understanding of certain terms used herein. The impeetigo encoding the therapeutic protein and albumin protein, once part 5 of the albumin fusion construct, may each be called “portion”, “region” or “moiety” of the albumin fusion construct.

The present invention relates generally to polynucleotides encoding albumin fusion proteins impettigo beta-interferon and their use in the treatment, prevention or amelioration of diseases or disorders. As used herein, “fusion protein alumina” refers to a protein formed by the fusion of 1 October albumin molecule to one molecule of a therapeutic protein.

A fusion protein albumin of the invention comprises a therapeutic protein and human serum albumin, which are associated with one another by genetic fusion i. Therapeutic protein and albumin protein, once part of the fusion construct 15 albumin, may each be called “portion”, “region” or impetivo of the fusion protein albumin p.

In a further preferred embodiment, a fusion protein of albumin of the invention is processed by a host and secreted into the culture medium surrounding cell.

Processing the fusion protein nascent albumin produced in the secretory pathways of the host used for expression may include, among others, 20 signal peptide cleavage, disulfide bond formation, proper folding, addition and processing of carbohydrates such as, for example, N-linked glycosylation and Oand specific proteolytic cleavages assembly multimeric proteins.

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A fusion protein albumin of the invention is preferably ipmetigo processed form.

In a more preferred embodiment, the “processed form of a fusion protein albumin” refers to a product of the fusion protein of albumin which has undergone excision of the signal peptide at N terminal, herein also called “mature fusion protein albumin”. Furthermore, it is lmpetigo to recover a fusion construct albumin given the deposit by techniques known in the art and are described elsewhere herein.

In one embodiment, the invention provides a polynucleotide encoding a fusion mipetigo comprising albumin, or alternatively consisting of, a therapeutic protein and a serum albumin protein. In a further embodiment 35, the invention provides an albumin fusion protein comprising, or alternatively consisting of, a Therapeutic protein and a serum albumin.

In a preferred embodiment, the invention provides an albumin fusion protein comprising, or alternatively consisting of, a therapeutic protein and a serum albumin protein encoded by a polynucleotide described in Table 2. The invention further encompasses polynucleotides encoding these albumin fusion proteins. The therapeutic protein portion of the albumin fusion protein is the mature portion of the therapeutic protein. As noted above, a polynucleotide of the invention encodes a protein comprising, or alternatively consisting of, a therapeutic protein and human serum albumin, which are associated with one another, preferably by genetic fusion.

As non-limiting example, a “therapeutic protein” is a protein that is useful for treating, preventing or ameliorating a disease, condition or disorder. As non-limiting example, a “therapeutic protein” may be impetito that binds specifically to a 55 particular cell type normal p. Specifically, a biological activity that is useful for treating, preventing or ameliorating a disease. Por ejemplo, se puede analizar un efecto deseable en un cultivo celular.

The therapeutic activity can be measured in vivo or in vitro.

ES2500918T3 – Albumin fusion proteins and beta interferon – Google Patents

Examples of assays include, among others, 10 described herein in the Examples section or column ” example activity assay ” column 3 Table 1. Corresponding to a therapeutic protein portion of a fusion protein of albumin of the invention, such as cell surface proteins and secretory therapeutic proteins often are modified by attachment of one or more oligosaccharide groups. The modification called glycosylation, can dramatically affect 15 to the physical properties of proteins and can be important in stability, secretion, and localization of proteins.

Glycosylation occurs at specific locations in the polypeptide structure. Typically, N-acetylneuraminic acid also known as sialic acid is the terminal residue of N-linked oligosaccharides attached to and O.

Variables such as protein structure and cell type influence the number and nature of the carbohydrate units within the chains at different glycosylation sites. Glycosylation isomers are also common at the same site within a given cell type.

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For example, several types of human interferon are glycosylated. The host cell into which they are expressed may modify the corresponding therapeutic proteins to the therapeutic protein portion of a fusion protein of albumin of the invention, in such a way that glycosylation at one or more sites is altered as a result of manipulation s of their nucleic acid sequence, or can be modified due to other conditions of their expression.

For example, can be produced isomers glycosylation 35 annulling or introducing glycosylation sites, for example by substitution or deletion of amino acid residues, such as substitution of glutamine for huemdo, or produced recombinant non-glycosylated proteins by expressing proteins in host cells not glycosylated, for example in E. These approaches are described in more detail below and are known in the art.

Therapeutic proteins, particularly those disclosed in Table 1, and nucleic acid sequences and amino acids are well known in impetiho art and available in public databases such as seoc for chemical abstraes p data.

X” 45 Table 2. X to derive the construct described in the same fila. X corresponds to a protein length but only a portion of that protein is used to generate the specific CID, it is impetig the skill in the art using techniques of molecular, such biology PCR, to amplify the specific fragment and clone it into the appropriate vector.

Table 1 provides a list of therapeutic proteins corresponding to a therapeutic protein portion of a fusion protein of albumin of the invention or a fusion protein of albumin encoded by a polynucleotide of the invention. The “therapeutic protein X” as used herein, can refer to a protein molecule individual therapeutic or entire group of therapeutic proteins associated with a given therapeutic protein molecule disclosed in this column. The column “biological activity” column 2 describes the biological activities associated with the therapeutic protein molecule.

Each of the references cited in the “activity test sample” column are incorporated herein by reference in its entirety, particularly with respect to the description of the assay respective activity described in the reference see section Procedures it, for example to analyze the activity 10 biological forth in “biological activity” in Table 1.

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The “ID Construction” column 5 column provides a link to an example of fusion construct albumin reported in Table 2 which encodes a fusion protein of 15 albumin comprising or alternatively, consisting of portion of the therapeutic protein X or fragment thereof reference.

Tabla 2 table 2. Table 2 provides the polynucleotide of the invention comprising, or alternatively consisting of, nucleic acid molecules encoding a fusion protein of albumin. The first column, “Fusion No.

Column 2, “Construction ID” provides a unique numeric identifier for the polynucleotide of the invention. The ID of the constructs can be used to identify polynucleotide encoding proteins albumin fusion comprising or alternatively consisting of, 5, a portion of the respective therapeutic protein to the therapeutic protein: X indicated in the corresponding row of the table 1, in which said construction ID indicated in column 5.

The fourth column of Table 2, “Description” provides an overview of a fusion construct 10 albumin and the fifth column, “Expression vector” the vector in which a polynucleotide comprising cloned or, alternatively it is consisting of a nucleic acid molecule encoding a fusion protein of albumin given. In the art vectors they are known and are commercially available or are described elsewhere. For example, as described in the Examples, one “expression cassette” comprising, or alternatively consisting of, one or more of 1 a polynucleotide encoding a fusion protein of albumin 15 given, 2 a leader sequence, 3 a promoter region and 4 a transcription terminator, may be assembled into a convenient cloning vector and then passed to a mammalian expression.

For expression in NS0 cells, an expression cassette comprising, or alternatively consisting of a nucleic acid molecule encoding a fusion protein of albumin in pEE12 cloned.

Y” provides the amino acid sequence of the full length albumin fusion protein of the invention. Y shows the unprocessed form of the protein albumin fusion encoded, in other words the SEQ ID NO and shows the signal sequence, in the portion of HSA, and a therapeutic portion, all it encoded by the concrete construction. Compositions comprising these two preferred embodiments, including pharmaceutical compositions are also preferred. X” provides the sequence of parental nucleic acid from which one can derive a polynucleotide encoding a therapeutic protein portion of a fusion protein of albumin given.

In one embodiment, the sequence parental nucleic acid from which one can derive a polynucleotide encoding a therapeutic protein portion of a fusion protein comprising albumin 40 wild gene sequence encoding a therapeutic protein shown in table 1.

This parent sequence may be a parental full length protein used to derive the concrete construction, the mature portion a parent protein, a variant or fragment of a wild-type protein, or an artificial sequence may be used to create the construction described. Z to determine which residues 50 amino acids of a protein albumin fusion encoded by a given construction are provided by the therapeutic protein. Z to derive the construct described in same row.

A” and “SEQ ID NO B”, respectively, are examples of primers used to generate a polynucleotide comprising or alternatively consisting of, a molecule of nucleic acid encoding the therapeutic protein portion of a fusion protein albumin given. X of the row as template DNA. PCR methods are well established in the art. Those skilled in the art could readily envision additional sequences useful primers. Oligonucleotide primers may be used in overlapping PCR reactions to generate mutations within a template DNA sequence.

In the art known PCR. Tabla 3 table 3. You may recover albumin fusion construct given the deposit by techniques known in the art and are described elsewhere herein see Example In a further embodiment of the invention an “expression cassette” comprising, or alternatively consisting of, one or more of 1 a polynucleotide encoding a fusion protein of albumin 15 given described, 2 a sequence leader, 3 a promoter region and 4 a transcription terminator can be moved or “subcloned” a vector to another.

Fragments to be subcloned can be generated by methods well known in the art such as, for example, PCR amplification p. Such functional activities include, among others, biological activity, antigenicity [ability to bind or compete with a polypeptide for binding to an anti-polypeptide antibody], the 25 immunogenicity ability to generate antibodies that bind to a polypeptide specific of the inventionability to form multimers with polypeptides of the invention and the ability to bind to a receptor or ligand for a polypeptide.

In case where there is dose dependent, need not be identical to the polypeptide, but substantially similar to the dose dependence in a given compared to the polypeptide of the present invention activity ie, the candidate polypeptide will exhibit greater activity or not more than about 25 times less, and preferably no more than about tenfold less activity, and most preferably, no more than about three times less activity 35 to the polypeptide of the present invention.

Se puede analizar la actividad funcional p.

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